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Molecular imaging and biology
Feb, 2024;26 (1): 148-161.

Reprogramming a Doxycycline-Inducible Gene Switch System for Bacteria-Mediated Cancer Therapy.

Hien Thi-Thu Ngo, Dinh-Huy Nguyen, Sung-Hwan You, Khuynh Van Nguyen, So-Young Kim, Yeongjin Hong, Jung-Joon Min
Author Information
  1. Hien Thi-Thu Ngo: Institute for Molecular Imaging and Theranostics, Chonnam National University Medical School, Gwangju, 61469, Republic of Korea.
  2. Dinh-Huy Nguyen: Institute for Molecular Imaging and Theranostics, Chonnam National University Medical School, Gwangju, 61469, Republic of Korea.
  3. Sung-Hwan You: Institute for Molecular Imaging and Theranostics, Chonnam National University Medical School, Gwangju, 61469, Republic of Korea.
  4. Khuynh Van Nguyen: Institute for Molecular Imaging and Theranostics, Chonnam National University Medical School, Gwangju, 61469, Republic of Korea.
  5. So-Young Kim: Institute for Molecular Imaging and Theranostics, Chonnam National University Medical School, Gwangju, 61469, Republic of Korea.
  6. Yeongjin Hong: CNCure Biotech, Hwasun, 58128, Republic of Korea. yjhong@jnu.ac.kr.
  7. Jung-Joon Min: Institute for Molecular Imaging and Theranostics, Chonnam National University Medical School, Gwangju, 61469, Republic of Korea. jjmin@jnu.ac.kr. ORCID
PMID: 38017353 DOI: 10.1007/s11307-023-01879-6

Abstract

PURPOSE: Attenuated Salmonella typhimurium is a potential biotherapeutic antitumor agent because it can colonize tumors and inhibit their growth. The present study aimed to develop a doxycycline (Doxy)-inducible gene switch system in attenuated S. typhimurium and assess its therapeutic efficacy in various tumor-bearing mice models.
PROCEDURES: A Doxy-inducible gene switch system comprising two plasmids was engineered to trigger the expression of cargo genes (Rluc8 and clyA). Attenuated S. typhimurium carrying Rluc8 were injected intravenously into BALB/c mice bearing CT26 tumors, and bioluminescence images were captured at specified intervals post-administration of doxycycline. The tumor-suppressive effects of bacteria carrying clyA were evaluated in BALB/c mice bearing CT26 tumors and in C57BL/6 mice bearing MC38 tumors.
RESULTS: Expression of the fimE gene, induced only in the presence of Doxy, triggered a unidirectional switch of the P promoter to induce expression of the cargo genes. The switch event was maintained over a long period of bacterial culture. After intravenous injection of transformed Salmonella into mice bearing CT26 tumors, the bacteria transformed with the Doxy-inducible gene switch system for Rluc8 targeted only tumor tissues and expressed the payloads 2 days after Doxy treatment. Notably, bacteria carrying the Doxy-inducible gene switch system for clyA effectively suppressed tumor growth and prolonged survival, even after just one Doxy induction.
CONCLUSIONS: These results suggest that attenuated S. typhimurium carrying this novel gene switch system elicited significant therapeutic effects through a single induction triggering and were a potential biotherapeutic agent for tumor therapy.

Keywords

DNA recombinase Doxycycline-inducible gene switch Salmonella-mediated cancer therapy Synthetic biology Tet promoter

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MeSH Term

Mice
Animals
Doxycycline
Mice, Inbred C57BL
Neoplasms
Plasmids
Bacteria

Chemicals

Doxycycline
Journal Article
Article has an altmetric score of 1

Word Cloud

Created with Highcharts 10.0.0switchgenetumorssystemmicetyphimuriumDoxycarryingbearingSDoxy-inducibleRluc8clyACT26bacteriatumorAttenuatedSalmonellapotentialbiotherapeuticagentgrowthdoxycyclineattenuatedtherapeuticexpressioncargogenesBALB/ceffectspromotertransformedinductiontherapyPURPOSE:antitumorcancolonizeinhibitpresentstudyaimeddevelop-inducibleassessefficacyvarioustumor-bearingmodelsPROCEDURES:comprisingtwoplasmidsengineeredtriggerinjectedintravenouslybioluminescenceimagescapturedspecifiedintervalspost-administrationtumor-suppressiveevaluatedC57BL/6MC38RESULTS:ExpressionfimEinducedpresencetriggeredunidirectionalPinduceeventmaintainedlongperiodbacterialcultureintravenousinjectiontargetedtissuesexpressedpayloads2 daystreatmentNotablyeffectivelysuppressedprolongedsurvivalevenjustoneCONCLUSIONS:resultssuggestnovelelicitedsignificantsingletriggeringReprogrammingDoxycycline-InducibleGeneSwitchSystemBacteria-MediatedCancerTherapyDNArecombinaseDoxycycline-inducibleSalmonella-mediatedcancerSyntheticbiologyTet

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